The Preliminary Research On Transfering Gmsultr1;2b Gene Into Soybean

Soybean[Glycine max (L.) Merr.] is a high-quality protein resources, and is one of the most important sources of plant protein for humans and animals. Soybean protein amino acid composition is complete, but low content of sulfur amino acids (Cys and Met), limits the nutritional value of soybean. And yet the plant started with sulfur assimilation synthesis of sulfur-containing amino acids. Sulfur transporter protein is necessary for the plant to absorb sulfate and the transport of sulfur. So sulfur transporters are important influence factors in the sulfur-containing amino acid metabolic pathways. In the former study of our lab, by GmSultrl;2b gene into tobacco and verify its function.The results showed that GmSultrl;2b is participation in SO42- absorption, and it could contribute to the assimilation of sulfur metabolism, improves sulfur amino acid content and yield of plants overexpressed GmSultr1;2b.Soybean storage protein gene expression is regulated by both transcriptional and post-transcriptional processes. RNA interference technology is a new gene disruption technology, that inhibits the expression of plant genes in the post-transcriptional level and regulate gene. Soybean 11S and 7S globulin was significantly negatively correlated, thus reducing 7S globulin content or increasing 11S globulin content could improve the nutritional quality and functional characteristics of soybean protein. The total protein and fat content of soybean will not be changed.This thesis mainly includes the following aspects:(1) We reconstructed the over expressing vector of GmSultrl;2b gene by double digestion method, and has been resistant to glufosinate-containing over-expression vector (pBA002-GmSultrl;2b). To get the higher expression soybean plants and improve soybean sulfur-containing amino acid content, this paper uses Agrobacterium-mediated genetic transformation of soybean cotyledon node. Also for the soybean genetic transformation system is optimized. Experiments finally got 234 soybean.transplants, Through a variety of post-detection methods five were identified as positive transgenic soybeans. The transformation receptors are all Jack, and with 21 seeds for to plants.(2) By T2 generation of pBl-Gm7S a positive transgenic soybean, moleculor detection and determination of amino acid content showed that most seeds sulfur-containing amino acids were increased. Selected the fine individual on-demand line, harvested seeds, measured its sulfur amino acids and found that the sulfur-containing amino acids of T3 generation seeds were increased too.(3) The advanced generation strains with RNAi-Gm7S a was selected by pedigree method. The seeds of the superior individual plants made the trip after harvest. Respectively by PCR and semi-quantitative analysis, then continuously measure the sulfur amino acids. Results showed that in soybean with low Gm7S a gene expression had a high sulfur amino acids score, and can be stably inherited.